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Tuesday, June 19, 2012

The signs are that the next RNAi Therapeutics
metabolic/cardiovascular disease candidate will be a dual-targeting one. While the initial attempts in this area were directed at specifically reducing the well-known cardiovascular risk factor LDL-cholesterol (preferred targets: ApoB and PCSK9), it has become quite tempting to
exploit the rare opportunity offered by RNAi Therapeutics to target multiple gene
targets with just one formulation to both broaden the therapeutic benefits
in patients that typically suffer from a plethora of metabolic dysfunctions
(obesity, insulin resistance, and hypercholesterolemia to name a few) and to
balance the adverse effects that may result from inhibiting certain targets.

Most notable among the latter is the liver
fat accumulation following ApoB knockdown.
Clinical studies with ISIS Pharmaceuticals’ antisense compound
mipomersen/KYNAMRO have clearly evidenced such liver fat accumulations which were often
accompanied by increases in liver enzymes, general indicators of liver
toxicity. These results further are corroborated
by similar clinical observations with the small molecule lomitapide by Aegerion
targeting microsomal triglyceride transfer protein (Mtp) which acts essentially
at the same stage as ApoB in packaging triglycerides and cholesterol for transport out of
liver cells into the circulation. Aegerion
obtained this drug candidate from BMS via UPenn as BMS did not want to further develop
this compound due to these safety risks.

Both mipomersen and lomipatide have completed phase III studies and new drug
applications for approval in the rare genetic disease homozygous familial hypercholesterolemia (hoFH), and in the case of mipo also for
severe heterozygous FH have been submitted to the FDA and EMA. In terms of therapeutic profile, mipomersen seems to have the edge as, being a phosphorothioate antisense compound, it preferentially accumulates in the liver. Consequently, it does not cause the side effects
resulting from the intestinal inhibition of this pathway that have been observed with small molecule lomatipe (note: SNALP-delivered RNAi Therapeutics should have similar benefits over small molecules). Moreover, mipomersen not only lowers LDLc, but also moderately reduces the
independent cardiovascular risk factor Lp(a).
Although not a prospective primary goal of mipomersen clinical development,
incidental positive findings like this one can go a long way in having regulators
take a benevolent look at drug candidates. This can be seen in the related
obesity space where one of the attractive benefits of Arena Pharmaceutical's lorcaserin is that it lowers blood glucose levels.

Obviously, there should be plenty of potential gene targets
involved in triglyceride synthesis and utilization/oxidation that could be exploited to
concomitantly lower triglyceride content in ApoB/Mtp-inhibited livers while maintaining LDLc-lowering.

Tep and colleagues from Merck published a paper on a study that tested whether an RNAi co-knockdown strategy could be implemented to alleviate the liver fat accumulations due to Mtp and ApoB inhibition. To be clear, Merck did not state that they have firm intentions of developing such a co-knockdown strategy, but nevertheless noted that such a strategy would have the advantage of not having to ‘add[ ] a novel compound on top of an approved drug’ and that
dual-targeting RNAi Therapeutics candidates are already in clinical
development, therefore paving the regulatory path (see ALN-VSP02, and
TKM-EBOLA).

In a first step the scientists confirmed the liver fat accumulation
following Mtp and ApoB siRNA knockdown.
Not only were they of similar magnitude, the
effects of the two knockdowns where essentially the same in almost every other
investigated regard. Notably, there was
no reduction in liver fat accumulation following prolonged siRNA treatment as
one might have expected based on claims by ISIS Pharmaceuticals of liver fat normalizations with time, but widespread
changes in the expression of lipid-related genes were nevertheless observed- this time
consistent with claims by ISIS Pharmaceuticals.

Among the genes that were downregulated following Mtp siRNA
treatment, presumably as a result of negative feedback, was DGAT2, a key enzyme in triglyceride synthesis that is also thought to represent an important
regulatory node in lipid metabolism (e.g. by promoting fatty acid oxidation). Reasoning that further reducing DGAT2 with
liposomally formulated siRNAs may lead to a measurable reduction in liver fat, they then co-formulated the Mtp siRNA with a DGAT2 siRNA and
injected them into mice. Indeed, this
resulted in not only the expected LDL-cholesterol reduction, but liver
triglyceride increases were mitigated.
According to data not shown, it was claimed that the same
beneficial effect could not be observed with an ApoB-DGAT2 siRNA combination, suggesting that Mtp may be the better target
for co-knockdown strategies.

It should be added, however, that the day 14 time-point data
these conclusions were based on were somewhat of an outlier as at this time the
co-formulation with DGAT2 siRNA reduced the LDLc-lowering potency of Mtp
knockdown. On the other hand, the
scientists report (also in data not shown) that they tested the
co-inhibition strategy using DNA-directed RNAi and thus validated this conclusion. Moreover, given the
multitude of genes involved in lipid metabolism, the Merck scientists stated
that the Mtp-DGAT2 co-knockdown is a proof-of-concept and that other targets besides DGAT2 are also being considered (especially Gpat1).

Given Tekmira’s interest in ApoB as a target and recent LNP
work by Alnylam and their PCSK9 collaborators from UT Southwestern (Horton,
Goldstein) on SCAP knockdown to alleviate hepatic steatosis and Alnylam's general interest in co-knockdown for metabolic
applications, Merck will not be alone in their endeavor of finding an RNAi
Therapeutic candidate that can do it all, LDLc lowering, triglyceride lowering,
and more. Given that SNALP technology would likely be used in such a clinical program, this could particularly benefit Tekmira.

4 comments:

Anonymous
said...

Merck also reported "AAV8-mediated Over-expression of Cynomolgus LCAT Raises HDL Cholesterol Levels in Cynomolgus Monkeys" which suggests they may be looking at a ddrnai/gene therapy approach delivered via an AAV vector.

Interesting one. All we have heard of ddRNAi of late from the media is delays to the Benitec pipeline,unlike the zippy but now defunct Nucleonics when they sped into their HBV trial.

What everyone has missed in the smoke of last couple of days however is City of Hope enrolling patients for next HIV trial using ddRNAi. Outcome of this trial could really crank things up. But listen to John Rossi, not me.

Single formulation with multiple targets knockdown strategy definitely is a unique and strong advantage for RNAi technology. RNAi technology certainly has broader reach than antibody based therapeutics. On top of those advantages, many undruggable targets could potentially be targeted with RNAi. We just need clinical proof.............

well the proof may not be far away. the small safety trial is seeing in one patient poliferation of HIV resistant T cells. That's 3.5years after infusion of the modified stem cells with the Pol III expression of shRNA trigger. The trial recruiting now increases to efficacy dose of modified stem cells..so if this comes good that'll be a pure play, black and white piece of clinical proof of RNAi. HIV one shot not as commercially attractive to big pharma obviously as say cholesterol, but proof none the less enough to get Merck moving on their prelim ddRNAi work. And Rossi mentioned influenza, HCV, and HBV as other diseases well in firing line of ddRNAi.

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